NMR IN BIOPHARMA

NMR IN BIOPHARMA



Abstract

The analysis of protein pharmaceuticals currently involves a complex series of chromatographic, electrophoresis, spectroscopic, immunological and biological measurements to unequivocally establish their identity, purity and integrity. In this review, I briefly consider the possibility that at least the functional identity and integrity of a protein drug might be established by a single analysis involving X-ray diffraction, NMR or mass spectrometry, or by a chromatographically based multi- detector system in which a number of critical parameters are essentially simultaneously determined. The use of a protein standard to obtain comparative measurements and new advances in the technology of each of these methods is emphasized. Advances in NMR spectroscopy and related computational methods continue at a rapid pace. In the past three years, the capability to make complete assignments of protein spectra has expanded from a limit of approximately 100 residues to a limit of possibly 400 residues via isotope-edited three- and four-dimensional methods. A current major obstacle to the implementation of these approaches is the frequent micro heterogeneity of protein preparations. The evolution of biological assays into measurements examining more defined intracellular signal transduction events or based on novel biosensors as well as the analysis of vaccines is also briefly discussed.

NMR IN BIOPHARMA

Introduction

Nuclear magnetic resonance (NMR) - the resonance absorption of electromagnetic energy of a substance containing nuclei with nonzero spin in an external magnetic field caused by the reorientation of magnetic moments of nuclei.

The phenomenon of magnetic resonance was discovered in 1,945 - 1946 years. The same nucleus of atoms in various environments in the molecule show different NMR signals. Unlike a NMR signal from the signal standard substance to determine the so-called chemical shift, which is due to the chemical structure of the studied material. In the methods of NMR there are many possibilities to determine the chemical structure of substances, conformation, mutual influence, the intra molecular transformation. The heart of the NMR spectrometer is a powerful magnet .In the experiment, the first implementation in practice Purcell, a sample placed in a glass ampoule with a diameter of 5 mm is between the poles of a strong electromagnet. Then, to improve the homogeneity of the magnetic field, the vial begins to rotate, and the magnetic field acting on it, gradually increase. The radiation source was used radio-frequency generator of high quality factor. Under the effect of increasing the magnetic field are beginning to resonate with the core, which is configured spectrometer. Frequency is increased to as long as it reaches a certain limit, above which the resonance is impossible. Since reaching the bridge currents are very small, removing one of the spectrum are not limited to, as do several dozen passes. All received signals are summed at the final schedule, the quality of which depends on the SNR device. In this method, the sample is exposed to RF energy constant frequency, while the strength of the magnetic field changes, so it is also called the method of continuous irradiation ...